Anti-Raf1 Rabbit Polyclonal Antibody for WB, IHC-P, IF/ICC, ELISA - P04049

Core Product Specifications and Parameters

Biological Background: Raf1 Function and Localization

• RAF1 (also known as c-Raf, Raf-1) is a serine/threonine-protein kinase that serves as a critical link between membrane-associated Ras GTPases and the MAPK/ERK cascade, regulating cell proliferation, differentiation, apoptosis, and survival.
• Activation of RAF1 initiates a phosphorylation cascade through MEK1/2 and ERK1/2, ultimately controlling transcription of genes involved in cell cycle progression and transformation.
• RAF1 phosphorylates multiple downstream targets including BAD (at Ser-75), adenylyl cyclases (ADCY2, ADCY5, ADCY6), PPP1R12A, TNNT2, and modulates NF-kB signaling, motility, apoptosis, proliferation, and angiogenesis.
• Subcellular localization: RAF1 is found in the cytoplasm, cell membrane, mitochondrion, and nucleus, with translocation to the mitochondrion mediating anti-apoptotic effects via BCL2 binding.
• Tissue specificity: In skeletal muscle, isoform 1 is more abundant than isoform 2.
• Post-translational modifications: RAF1 undergoes phosphorylation at Ser-338 and Ser-339 by PAK1, and is subject to methylation, metal binding (zinc), and alternative splicing, giving rise to multiple isoforms.
• Associated with diseases: Mutations and dysregulation of RAF1 are linked to cardiomyopathies (CMD1NN) and deafness, underscoring its role in diverse physiological processes.
• Key biological roles: Acts as a proto-oncogene, regulating wound healing, epithelial cell transformation, and suppression of apoptosis.

Experimental Guidance and Technical Tips

• For Western blotting, the antibody detects a band at ~73 kDa. Positive controls such as HeLa, HEK293, or Jurkat cell lysates are recommended.
• In immunohistochemistry, antigen retrieval may be required; consider testing both heat-induced and enzymatic methods on paraffin-embedded tissues.
• For immunofluorescence, include Triton X-100 permeabilization and use appropriate secondary antibodies; note that Raf1 localizes to the cytoplasm, membrane, and nucleus.
• The immunogen spans amino acids 190-350, which may recognize multiple Raf1 isoforms; validation in relevant species (human, mouse, rat) is recommended.
• As a polyclonal antibody, lot-to-lot variability may occur; verify performance in the intended assay with appropriate controls.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio is a one-stop sourcing partner for IVD raw materials, supporting diagnostic developers from concept to clinic with validated antibody pairs, optimized monoclonal/polyclonal antibodies, bulk ancillary reagents, and rare target sourcing. For researchers and manufacturers focusing on the MAPK signaling pathway and oncology biomarkers, we offer this anti-Raf1 polyclonal antibody alongside a comprehensive portfolio of related raw materials to accelerate your assay development and clinical translation.